D-半乳糖诱导间充质干细胞衰老的研究  

Study on D-galactose-induced Senescence of Mesenchymal Stem Cells

作　　者：杨孟恺[1] 易诚青[1] Yang Mengkai;Yi Chengqing(Department of Orthopedics,Shanghai General Hospital Affiliated to Shanghai Jiaotong University,Shanghai 200080,Chin)

机构地区：[1]上海交通大学附属第一人民医院骨科,200080

出　　处：《医学研究杂志》2018年第7期31-36,共6页Journal of Medical Research

基　　金：国家自然科学基金资助项目(81371979)

摘　　要：目的利用D-半乳糖诱导建立间充质干细胞(mesenchymal stem cells,MSCs)体外衰老模型,并探讨其潜在机制。方法以正常培养的MSCs做对照组,以分别用1、10、100g/L D-半乳糖诱导的MSCs作为处理组,培养48h后,采用免疫蛋白印迹法(Western blot)检测细胞内AKT与mTOR蛋白磷酸化水平以及衰老相关蛋白p16Ink4a的变化。同时使用β-半乳糖苷酶试剂盒检测MSCs中β-半乳糖苷酶(SA-β-gal)表达量的变化。最后用AKT抑制剂预处理D-半乳糖诱导组以明确AKT/mTOR信号通路在MSCs衰老中的作用。结果与对照组相比,10g/L D-半乳糖作用48h后MSCs中β-半乳糖苷酶表达和衰老相关蛋白p16Ink4a含量明显增加,同时p-AKT与p-mTOR含量也较对照组增加,加入AKT抑制剂后,衰老组β-半乳糖苷酶表达和衰老相关蛋白p16Ink4a含量皆有所下降。结论 D-半乳糖能成功诱导构建MSCs体外衰老模型,AKT/mTOR信号通路参与了这一过程。Objective To establish the aging model of mesenchymal stem cells （MSCs） induced by D-galactose （D-gal） and investigate its underlying mechanism. Methods The MSCs were divided into control group and D-gal treatment group. Control group was normal cultured MSCs. D-gal treatment group was the control group treated with different concentrations of D-gal （1, 10, 100g/L）. All of them were treated for 48h. The expression of p16Ink4a protein, p-AKT, and p-mTOR were detected by Western blot. At the same time, the SA-β-galactose （SA-β-gal） staining was used to examined the senescence-associated changes. At last, AKT inhibitor was added in the D-gal treatment group to clarify the role of AKT/mTOR signaling pathway in MSCs aging. Results Compared with the control group, the quantity of SA-β-gal positive cells and the expression of p16Ink4a in MSCs were significantly increased in 10g/L D-gal treatment group. Meanwhile the expression of p-AKT and p-mTOR also increased compared with the control group. After adding in AKT inhibitor, the quantity of SA-β-gal positive cells in D-gal treatment group and the expression of p16Ink4a were decreased, too. Conclusion D-gal can successfully establish the aging MSCs model in vitro and AKT/mTOR signaling pathway is involved in this process.

关 键 词：D-半乳糖 信号通路 间充质干细胞 衰老 

分 类 号：R3[医药卫生—基础医学]